LOCUS       sequence-463312-        6884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6884)
  AUTHORS   Lopez Martinez D, Todorovski I, Noe Gonzalez M, Rusimbi C, Blears D,
            Khallou N, Han Z, Dirac-Svejstrup AB, Svejstrup JQ
  TITLE     PAF1C-mediated activation of CDK12/13 kinase activity is critical 
            for CTD phosphorylation and transcript elongation.
  JOURNAL   Mol Cell. 2025 May 15;85(10):1952-1967.e8. doi: 
            10.1016/j.molcel.2025.04.012. Epub 2025 May 1.
  PUBMED    40315851
REFERENCE   2  (bases 1 to 6884)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2025.04.012"; journalName: "Mol Cell"; date: 
            "2025-05-15- 15"; volume: "85"; issue: "10"; pages: "1952-1967.e8"
COMMENT     Sequence Label: pLIB-LEO1
FEATURES             Location/Qualifiers
     source          1..6884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  541..765
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(899..916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1070..1658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1150..1169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1829..2620,2621..2689))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2452..2471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2690..2794)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2820..3274)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2957..2978)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3168..3187
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  3458..3623
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    3733..3867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3757..3776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3811..3830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6040..6131)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(6098..6115)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    6179..6212
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(6608..6613,6614..6630,6631..6636)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(6825..6884,1..474)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ggctcaagta tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt caaatccatg
       61 cgggctgctc ttgatctttt cggtcgtgag ttcggagacg tagccaccta ctcccaacat
      121 cagccggact ccgattacct cgggaacttg ctccgtagta agacattcat cgcgcttgct
      181 gccttcgacc aagaagcggt tgttggcgct ctcgcggctt acgttctgcc caagtttgag
      241 cagccgcgta gtgagatcta tatctatgat ctcgcagtct ccggcgagca ccggaggcag
      301 ggcattgcca ccgcgctcat caatctcctc aagcatgagg ccaacgcgct tggtgcttat
      361 gtgatctacg tgcaagcaga ttacggtgac gatcccgcag tggctctcta tacaaagttg
      421 ggcatacggg aagaagtgat gcactttgat atcgacccaa gtaccgccac ctaacaattc
      481 gttcaagccg agatcggctt cccggccgcg gagttgttcg gtaaattgtc acaacgccgc
      541 gaatatagtc tttaccatgc ccttggccac gcccctcttt aatacgacgg gcaatttgca
      601 cttcagaaaa tgaagagttt gctttagcca taacaaaagt ccagtatgct ttttcacagc
      661 ataactggac tgatttcagt ttacaactat tctgtctagt ttaagacttt attgtcatag
      721 tttagatcta ttttgttcag tttaagactt tattgtccgc ccacacccgc ttacgcaggg
      781 catccattta ttactcaacc gtaaccgatt ttgccaggtt acgcggctgg tctatgcggt
      841 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
      901 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      961 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1021 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1081 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1141 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1201 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1261 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1321 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1381 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1441 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1501 acactagaag aatagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1561 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1621 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1681 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1741 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1801 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1861 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1921 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1981 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2041 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2101 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2161 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2221 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2281 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2341 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2401 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2461 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2521 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2581 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2641 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2701 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2761 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     2821 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2881 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2941 agggttgagt gttgttccag tttggaacaa gagtccacta ttaagaacgt ggactccaac
     3001 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3061 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3121 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3181 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3241 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcaaata
     3301 agcgttgata ttcagtcaat tacaaacatt aataacgaag agatgacaga aaaattttca
     3361 ttctgtgaca gagaaaaagt agccgaagat gacggtttgt cacatggagt tggcaggatg
     3421 tttgattaaa aacataacag gaagaaaaat gccccgctgt gggcggacaa aatagttggg
     3481 aactgggagg ggtggaaatg gagtttttaa ggattattta gggaagagtg acaaaataga
     3541 tgggaactgg gtgtagcgtc gtaagctaat acgaaaatta aaaatgacaa aatagtttgg
     3601 aactagattt cacttatctg gttcggatct cctaggggtt atgatagtta ttgctcagcg
     3661 gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc ttctaggctc
     3721 aagcagtgat cagatccaga catgataaga tacattgatg agtttggaca aaccacaact
     3781 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3841 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3901 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     3961 gctgattatg atcctctagt acttctcgac aagctttcaa tcatcatctt cttcctcttc
     4021 atcgctgatc acatacttct tatgcttttt atttgcttta tcatcatctt ctgcttttct
     4081 ctttccggaa ggttcacctt cctcatcact ggtaagtttc tttgctttga gtaatctttg
     4141 agctttatct tcttctgatc cctcatcact gtctgatgaa tagattctgg ctcgttcctc
     4201 tcgaatgccc cctttatatc ggtttttaat ggcagccaag ctgatggact cctcgccttc
     4261 ctcctcctca tcgtatcgat caggttccag gtaactggcg ctcagccccc gctggtgctg
     4321 tttctctctc attcggcgct gctgagattc cctacgtatg gaagccctca aacgttcttc
     4381 ttctttctta atcatttctg tgcgttggca ttcaggatca cgaccagcca ttggcaagat
     4441 tctaatcttc tgtgtctttg aacacctatc tgcaagtgac agagtcatct ttctatgtgt
     4501 ggcactgtcc gtagagtgag gtctgaaggt gagtttcgtt ttaaagactg cttgtccctg
     4561 tagaccagta ccttgtctta taaaaagatg attgtggtcg ccctgcagtg gggctttgta
     4621 cacatcaaac acttcattgc ctaaatgcag ggacatgctt ccatctgacc acttgactat
     4681 ccgagcattg ctttctttaa tttcatttcc ttcttcatct cggcgtatcc tccatcttat
     4741 agtattttct acctttaatt ttaacctggt tctaccttct tcatccagca tttcttcatc
     4801 ttcaaattca tcttcataat actgaggatc aaaaggtctg ggctctacac tgagaaagtt
     4861 gggcagttta acaaaatata agtcgtttcc taaatcagtg tttactttgg gtatttctac
     4921 ttctattctg gtctcaggaa ttggctcctc ttcctgttga tcctgaggca atccattttc
     4981 atcaacaggc tgtcctggag taggtggttt gtcttctcca tcactccctg aagagatatc
     5041 atctgcacct ccaaataaat ccatggttcc actattatct tttggcacct cagtgtcact
     5101 atccgcttct gaatcagatg caatcgcatt cttgcgtttc attcgtaaaa cttcatcttc
     5161 actatcactg cctcttgcag attctgattt atgatcctgt tcctcttcat catctgaatg
     5221 cctgtgttct tcatctgagg cctgtggcct ttcatcatca gaattttgca ttttttcttc
     5281 atcagacagc tgtggttgtt cttcatcatc agaattctgt ttctcatcat cattatcaga
     5341 agctaccggc cgttcatcat cagaattagc cttttcctcc tcagatagct gttgtctctc
     5401 atcatcggaa agctgaggcc tctcctcatc atctgtgttc tgcattttct catcatcgtc
     5461 agaattctgc agcttatctt catcagatcc ttgtgccctc tcctcatcat cagaattttg
     5521 tatcttttca tcatctgact ggtcactttt atcttctctg ccccattttt catcatctga
     5581 atgtgctttt tcagaacctt ctgcttctga atgatggctc cctccatccg atctatgacc
     5641 ttcgtcttca tcatcattag gggcttctga tccactgtgc tgatctacat ctgaggggtc
     5701 attgtcctca tggtcagaac gctcagaagc ttctgatcta ttgtctgatc tttcagagtg
     5761 attatcacta ccactatgat gtgaagctcc ctcgtcctca ctgtcatctc caaacagttc
     5821 cttattactt ggttgtcctg aatcacctct ttcatcctga tcactttcac ttccagaggc
     5881 attactgcca gaggcagcat tctcttgatc agaatctgag tcagatccag aatcagaatc
     5941 tttacgctca gcttcgctgt cggcgtcgct cccgaagaga tcctccatat ccgccatgga
     6001 tccgcgcccg atggtgggac ggtatgaata atccggaata tttataggtt tttttattac
     6061 aaaactgtta cgaaaacagt aaaatactta tttatttgcg agatggttat cattttaatt
     6121 atctccatga tctattaata ttccggagta ggtcgcgaat cgatactagt aaaaccagat
     6181 aacttcgtat aatgtatgct atacgaagtt atggtaccgc ggccgcgtag aggatctgtt
     6241 gatcagcagt tcaacctgtt gatagtacaa acaaagctgt agccaaccac tagaactata
     6301 gctagagtcc tgggcgaaca aacgatgctc gccttccaga aaaccgagga tgcgaaccac
     6361 ttcatccggg gtcagcacca ccggcaagcg ccgcgacggc cgaggtctac cgatctcctg
     6421 aagcaagggc agatccgtgc acagcacctt gccgtagaag aacagcaagg ccgccaatgc
     6481 ctgacgatgc gtggagaccg aaaccttgcg ctcgttcgcc agccaggaca gaaatgcctc
     6541 gacttcgctg ctgcccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcaca
     6601 aacccagttg acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc
     6661 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     6721 catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc
     6781 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca
     6841 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggt
//
